Abstract
Recombinant protein expression in Escherichia coli represents a cornerstone of the biotechnology enterprise. While cytoplasmic expression in this host has received the most attention, achieving substantial yields of correctly folded proteins in this compartment can sometimes be met with difficulties. These issues can often be overcome by targeting protein expression to extracytoplasmic compartments (e.g., membrane, periplasm) or to the culture medium. This chapter discusses various strategies for exporting proteins out of the cytoplasm as well as tools for monitoring and optimizing these different export mechanisms.
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Acknowledgment
J.T.B. gratefully acknowledges NSF GK12 fellowship support (DGE-1045513) and D.W. gratefully acknowledges the Royal Thai Government for fellowship support. This work was supported by DOE Great Lakes Bioenergy Research Center (GLBRC) Project 3.2.8, USDA NIFA Award # 2009–02202, and NIH Award # DA031409.
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Boock, J.T., Waraho-Zhmayev, D., Mizrachi, D., DeLisa, M.P. (2015). Beyond the Cytoplasm of Escherichia coli: Localizing Recombinant Proteins Where You Want Them. In: García-Fruitós, E. (eds) Insoluble Proteins. Methods in Molecular Biology, vol 1258. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2205-5_5
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DOI: https://doi.org/10.1007/978-1-4939-2205-5_5
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